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Rabies virus production in high vero cell density cultures on macroporous microcarriers
A Y Yokomizo1, M M Antoniazzi, P L Galdino
1Laboratório de Imunologia Viral, Instituto Butantan, 05503-900 São Paulo, Brasil. grugel@butantan.gov.br
Biotechnology and Bioengineering
|February 5, 2004
Summary
Porous microcarriers (MCs) like Cytopore and Cultispher G significantly enhance rabies virus production in Vero cells compared to solid MCs. These supports offer higher cell densities and virus titers, proving useful for rabies antigen production.
Area of Science:
- Biotechnology
- Virology
- Cell Culture Technology
Background:
- Rabies virus antigen production is crucial for vaccine development.
- Optimizing cell culture conditions, specifically microcarrier selection, can improve virus yield.
- Porous microcarriers offer increased surface area for cell attachment and growth compared to nonporous alternatives.
Purpose of the Study:
- To investigate rabies virus multiplication in Vero cell cultures using porous (Cytopore, Cultispher G) and solid (Cytodex 1) microcarriers (MCs).
- To compare cell densities, cell concentration increases, and rabies virus titers achieved on different types of MCs.
- To evaluate the impact of MC concentration on cell performance and virus production.
Main Methods:
- Vero cells were cultured on porous (cellulose-Cytopore, gelatin-Cultispher G) and solid (DEAE-Cytodex 1) microcarriers at consistent concentrations.
- Cell densities, cell concentration fold-increase, and rabies virus titers (FFD50/mL) were measured.
- Cell attachment rates, cell growth kinetics, and cell densities were analyzed.
- Microscopy (optical, scanning, transmission electron) was used to assess MC structure and cell distribution.
Main Results:
- Porous MCs (Cytopore, Cultispher G) achieved nearly double the cell densities compared to solid MCs (Cytodex 1).
- Cytopore and Cultispher G MCs showed significantly higher fold-increases in cell concentration and rabies virus titers than Cytodex 1.
- Higher MC concentrations led to reduced virus production and MC-cell occupation.
- Cell attachment was fastest on Cytopore and Cytodex 1 MCs, with similar exponential cell growth rates observed on all MC types.
Conclusions:
- Porous microcarriers, particularly Cytopore and Cultispher G, are effective supports for enhancing rabies virus antigen production in Vero cell cultures.
- Uniform cell distribution within porous MCs contributes to efficient virus synthesis.
- The findings highlight the potential for optimizing microcarrier-based cell culture systems for improved rabies virus production.